Antioxidant treatment ameliorates diabetes-induced dysfunction of the vas deferens in a rat model.

Diabetes mellitus (DM) affects the male ejaculatory function. This study was designed to evaluate the role of oxidative stress in the development of diabetes-induced dysfunction of vas deferens (VD) in the rat. DM was induced by streptozotocin in 40 male Wistar rats. Subsequently, the diabetic animals were divided into three groups: DM group, DM + Eda group and DM + Tau group. These groups were administered saline, edaravone and taurine, respectively, daily for 4 weeks. Another group of ten rats served as a control group. DM was diagnosed in the 40 streptozotocin-injected rats. DM significantly reduced the VD weight. Additionally, DM induced in vitro VD hypercontractility, VD histological abnormalities and increased the serum and VD tissue concentration of malondialdehyde. VD immunohistochemistry revealed overexpression of three markers of oxidative stress. DM significantly reduced serum testosterone levels. No live birth was documented in all DM rats in mating experiments. Antioxidants significantly improved all the aforementioned parameters, except the testosterone levels. This study indicates a deleterious impact of DM-induced oxidative stress on VD histological and functional features. Antioxidant treatment may provide an adjunct tool to alleviate ejaculatory disorders for male patients with type 1 diabetes.

Post-fertilization effect of bilateral primary testicular damage induced by unilateral cryptorchidism in the rat model.

Cryptorchidism, a common anomaly of the male genitalia, affects 2-4% of male infants. The post-fertilization effects of unilateral cryptorchidism model in the rat and the effects of antioxidant treatment were investigated. Six-week-old male Wistar rats were randomly separated into four groups. Unilateral cryptorchidism was induced in the right testis of three groups. One group was treated with saline intraperitoneally (i.p.) (Crypto), one group was treated with taurine (500 mg/kg, i.p.; Tau), and another group was treated with sivelestat (15 mg/kg i.p.; Siv). The control group was treated with saline i.p. The treatment was daily for 8 weeks. Five days before sacrifice, mating studies were performed. Body, testicular, and epididymal weights were recorded. Malondialdehyde (MDA) levels in the seminal vesicular fluid (SVF) were measured. Testicular levels of MDA and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were determined bilaterally. TUNEL assay was used to examine DNA fragmentation bilaterally. Histological examination and the Johnsen score were used to evaluate morphological testicular alterations. The Crypto group demonstrated significantly lower right testicular and epididymal weights, significantly increased SVF-MDA levels, testicular MDA and 8-OHdG levels, and the apoptotic score bilaterally compared to the controls. Furthermore, histological evaluation revealed significantly reduced spermatogenesis and mild injury to the cryptorchid testes compared to the control. Treatment with both taurine and sivelestat significantly reduced SVF-MDA levels, testicular MDA, 8-OHdG, and apoptosis bilaterally compared to the Crypto group. Antioxidant treatment was unable to ameliorate spermatogenesis. Newborns delivered by females that mated with Crypto-males had significantly lower body weight compared with the respective animals from the control, Tau and Siv groups. The present study demonstrated that unilateral cryptorchidism-induced testicular damage can significantly affect the contralateral testis as well having further deleterious post-fertilization effect on the development of newborns. Treatment with antioxidants can partially improve the testicular damage bilaterally with beneficial effects for the newborns.

Experience with injections of botulinum toxin type A into the detrusor muscle.

INTRODUCTION: We evaluated the effects of intra-vesical injection of botulinum toxin type A in the detrusor muscle in patients with neurogenic overactive bladder (OAB), patients with non-neurogenic overactive bladder and patients with interstitial cystitis. MATERIALS AND METHODS: Between January 2003 and December 2006 we treated 30 patients with 100 I. U. to 300 I. U. of botulinum toxin A in the detrusor muscle. Patients were clinically and urodynamically followed up for 4, 12 and 36 weeks thereafter. RESULTS: Neurogenic overactive bladder: of the 19 injected doses, 18 (94.7%) in 7 patients were judged as effective, and 1 (5.2%) of 200 U of BTX-A was judged as ineffective. Mean bladder volume increased from 137 to 396 ml. Non-neurogenic overactive bladder: of 7 injections, 6 (85.7%) were judged effective in 5 patients. Mean bladder volume increased from 149 to 322 ml. Interstitial cystitis: in all 4 patients the treatments were deemed ineffective. CONCLUSIONS: Injecting 300 units of BTX-A into 30 sites in the muscle located in the body of the bladder region is effective for neurogenic bladder patients with intermittent catheterization who have urge and reflective types of incontinence. Injections of 100 and 200 units of BTX-A to treat non-neurogenic overactive bladder with urinary incontinence provided together without retention. The optimal dose of BTX-A requires further investigation. Injection with 200 units of BTX-A was not useful against interstitial cystitis. None of the patients developed any adverse effects after injecting the bladder wall with BTX-A.

Bond durability of composite luting agents to ceramic when exposed to long-term thermocycling.

PURPOSE: This study evaluated the effect of thermocycling on the microtensile bond strength of four adhesive luting agents to GN-I CAD-CAM ceramic. The hypothesis tested was that thermocycling did not affect bonding effectiveness, irrespective of the luting agents used. MATERIALS AND METHODS: Ceramic specimens of two different sizes (6x8x3 mm; 13x8x4 nm) were fabricated from GN-I CAD-CAM ceramic blocks (GC) using a low-speed diamond saw. Two different sized porcelain discs were bonded with one of the four composite luting agents (Linkmax [LM], Panavia [PN], RelyX Unicem [UN] and Variolink II [VL]) according to the manufacturer's instructions. The specimens were stored for 24 hours in distilled water at 37 degrees C and subjected to 0; 10,000; 20,000 and 40,000 thermocycles prior to microTBS testing. Two-way analysis of variance was used to test the influence of luting cement, thermocycling and interaction between both (p < 0.05). The Tukey HSD test determined statistical differences in microTBS for each luting composite between the different thermocycling conditions (p < 0.05). The mode of failure was determined at a magnification of 50x using a stereomicroscope (Wild M5A). RESULTS: Two-way ANOVA revealed that microtensile bond strength was affected by the luting cement, thermocycling and a combination of both. No difference in bond strength between Linkmax, Panavia F and Variolink II was noticed after 24 hours of water storage (LM: 47.6 MPa; PN: 41 MPa; VL: 36 MPa). RelyX Unicem scored significantly lower than Linkmax and Panavia F (UN: 24.2 MPa). The influence of thermocycling on bond strength was different for the four luting cements. Using Variolink II, the bond strength remained stable after 40,000 thermocycles (43.6 MPa). Linkmax showed a significant decrease in bond strength after 10,000 (26 MPa) and 40,000 thermocycles (14.8 MPa). Panavia F and RelyX Unicem were the most negatively influenced, as all specimens failed prior to testing (pre-testing failures) when the specimens were thermocycled 10,000 and 20,000 times or longer, respectively. Regarding the failure mode, there was a correlation between bond strength and type of failure. Initially, a combination of adhesive and mixed adhesive-cohesive failures was noticed. The percentage of adhesive failures increased, together with a decrease in bond strength. CONCLUSION: It was concluded that there were significant differences among the four resin composite cements in terms of their bonding effectiveness to CAD-CAM ceramic after thermocycling. The varying degrees of bonding effectiveness of these adhesive luting agents highlight the need for material specifications.

Bonding effectiveness of adhesive luting agents to enamel and dentin.

OBJECTIVES: The bonding effectiveness of five adhesive luting agents to enamel and dentin using different application procedures was determined using a micro-tensile bond strength protocol (microTBS). METHODS: Enamel/dentin surfaces of human third molars were flattened using a high-speed diamond bur. Composite resin blocks (Paradigm, 3M ESPE) were luted using either Linkmax (LM; GC), Nexus 2 (NX; Kerr), Panavia F (PN; Kuraray), RelyX Unicem (UN; 3M ESPE) or Variolink II (VL; Ivoclar-Vivadent), strictly following manufacturers' instructions. For some luting agents, modified application procedures were also tested, resulting in four other experimental groups: Prompt L-Pop+RelyX Unicem (PLP+UN; 3M ESPE), Scotchbond Etchant+RelyX Unicem (SE+UN; 3M ESPE), Optibond Solo Plus Activator+Nexus 2 (ACT+NX; Kerr) and K-Etchant gel+Panavia-F (KE+P; Kuraray). The experimental groups were classified according to the adhesive approach in self-adhesive (UN), etch-and-rinse (ACT+NX, NX, KE+P, SE+UN and VL when bonded to enamel) and self-etch adhesive luting agents (LM, PLP+UN, PN and VL when bonded to dentin). The specimens were stored for 24h in distilled water at 37 degrees C prior to microTBS testing. The Kruskal-Wallis test was used to determine pairwise statistical differences (p<0.05) in microTBS between the experimental groups. RESULTS: When bonded to enamel, ACT+NX (15 MPa) and UN (19.6 MPa) scored significantly lower than VL (49.3 MPa), LM (49.2 MPa), PN (35.4 MPa) and SE+UN (35.2 MPa), while PLP+UN (23.5 MPa) showed a significantly lower microTBS than VL (49.3 MPa) and LM (49.2 MPa). No significant differences were noticed between VL (49.3 MPa), LM (49.2 MPa), NX (37.9 MPa), KE+PN (38.8 MPa), PN (35.4 MPa) and SE+UN (35.2 MPa). Regarding the bonding effectiveness to dentin, all luting agents bonded equally effectively (UN: 15.9 MPa; LM: 15.4 MPa; PN: 17.5 MPa; NX: 22.3 MPa), except VL (1.1 MPa), SE+UN (5.9 MPa) and ACT+NX (13.2 MPa). VL revealed an exceptionally high number of pre-testing failures, most likely due to a combined effect of not having cured the adhesive separately and an insufficiently light-cured luting agent. SIGNIFICANCE: Following a correct application procedure, the etch-and-rinse, self-etch and self-adhesive luting agents are equally effective in bonding to enamel and dentin. Several factors negatively influenced bond strength such as bonding RelyX Unicem to enamel without prior phosphoric acid etching; no separate light-curing of a light-polymerizable adhesive prior to cementation, use of a light-polymerizing adhesive converted into a dual-polymerizing adhesive, and use of a dual-cure luting agent with a low auto-polymerizable potential.

Effects of ceramic surface treatments on the bond strength of an adhesive luting agent to CAD-CAM ceramic.

OBJECTIVES: The objective of this study was to evaluate the effect of different surface treatments on the micro-tensile bond strength (microTBS) of an adhesive luting agent to CAD-CAM ceramic. The hypothesis tested was that neither of the surface treatments would produce higher bond strength of the adhesive luting agent to CAD-CAM ceramic. METHODS: Ceramic specimens of two different sizes (6 mm x 8 mm x 3 mm; 13 mm x 8 mm x 4 mm) were fabricated from ProCAD ceramic blocs (Ivoclar-Vivadent) with a low-speed diamond saw. The ceramic blocks were divided into seven groups and submitted to the following surface treatments: group 1: no treatment; group 2: etching with 37% H(3)PO(4); group 3: etching with 37% H(3)PO(4)+silane; group 4: etching with 37% H(3)PO(4)+silane+adhesive resin; group 5: etching with 4.9% HF acid; group 6: etching with 4.9% HF acid+silane; group 7: etching with HF acid+silane+adhesive resin. After surface treatment, two differently sized porcelain disks were bonded together with a composite luting agent (Variolink II, Ivoclar-Vivadent). The specimens were stored for 24h in distilled water at 37 degrees C prior to microTBS testing. One-way analysis of variance was used to test the influence of surface treatment and Scheffe multiple comparisons test determined pair-wise statistical differences (p<0.05) in microTBS between the experimental groups. RESULTS: The mean microTBSs (standard deviation) are: group 1: 12.8 (+/-4.6)MPa; group 2: 19.1 (+/-5.0)MPa; group 3: 27.4 (+/-11.1)MPa; group 4: 34.0 (+/-8.9)MPa; group 5: 37.6 (+/-8.4) MPa; group 6: 34.6 (+/-12.8)MPa; group 7: 34.5 (+/-5.1)MPa. Statistical significant differences were found between group 1 and groups 3-7, and between group 2 and groups 4-7. All specimens of groups 1-4 exhibited adhesive failures, while a combination of adhesive and mixed (adhesive and cohesive) failures was observed in the specimens of groups 5-7. CONCLUSIONS: The results show that surface treatment is important to bond to ceramic and suggests that etching is needed preferably with hydrofluoric acid than with phosphoric acid.

Active surveillance for acute flaccid paralysis in poliomyelitis high-risk areas in southern China.

OBJECTIVE: On 29 October 2000 poliomyelitis was officially declared to have been eradicated from the Western Pacific Region. This article describes the results of surveillance for cases of acute flaccid paralysis (AFP) in China during the final phase of the eradication effort. METHODS: We conducted hospital-based active surveillance in high-risk areas for poliomyelitis in 5 provinces of southern-China (Sichuan, Yunnan, Guizhou, Guangxi and Jiangxi) between 1995 and 1997 to determine the adequacy of reporting and laboratory diagnosis of cases of AFP. FINDINGS: A total of 1069 AFP cases occurring since 1993 were identified in 311 hospital visits. Less than 50% of AFP cases occurring in 1993 and 1994 had been reported by AFP surveillance, and laboratory diagnosis had been carried out on only a small proportion of these. However, improved cooperation between hospital sectors increased the rate of case reporting and laboratory diagnosis to 85% and 78%, respectively, in 1997. Despite this overall improvement, these two indicators were approximately 10-20% lower in Yunnan Province. Epidemiological analysis revealed that cases of clinical poliomyelitis accounted for as much as one-third of all AFP in 1993 and that some of these cases were clustered. Clusters were rarely observed after 1994. Active surveillance in the China-Myanmar border areas of Yunnan over 1995-96 detected 9 cross-border cases of clinical poliomyelitis, including 2 of wild poliomyelitis. Import of poliomyelitis was thus considered to have occurred frequently until 1996 in this border area of Yunnan. These data were important for the outbreak response immunization carried out in 1996 in the border prefectures of Yunnan. CONCLUSION: Our investigation confirmed a high level of AFP surveillance in poliomyelitis high-risk areas of the five provinces and provided valuable information on the interruption of wild poliovirus circulation in southern China that will be of use to countries in other regions that have yet to eradicate poliomyelitis.

Transcriptional activation function of zinc finger protein TIS11 and its negative regulation by phorbol ester.

TIS11, a CCCH zinc finger protein, is one of the typical growth factor-inducible nuclear proteins. We found that TIS11 possesses the potential to activate transcription when fused to the GAL4 DNA binding domain and transiently cotransfected into rat pheochromocytoma PC12 cells along with a GAL4-responsive luciferase reporter gene. The study with deletion mutants of TIS11 revealed that the major transactivation region is located at the N-terminal 101 amino acid residues and that the remaining central and C-terminal region had a moderate transactivational activity. In addition, the transactivational activity of TIS11 was found to be significantly reduced by treating the transfectants with phorbol 12-myristate 13-acetate (PMA). PMA-induced inactivation of TIS11 was blocked by calphostin C, a protein kinase C inhibitor, and PD98059, a mitogen-activated protein (MAP) kinase kinase inhibitor. These results suggested that TIS11 functions as a positive transcriptional regulator and that the protein kinase C/MAP kinase signaling cascade is involved in negative regulation of TIS11 by PMA.

Identification of the essential cysteinyl residue located in the active site of human phenylethanolamine N-methyltransferase.

Phenylethanolamine N-methyltransferase (PNMT) catalyzes the production of epinephrine from norepinephrine using S-adenosyl-L-methionine as a methyl donor. Previous studies of chemical modification of the PNMT with reagents specific to Cys residues showed that the enzyme contains a Cys residue essential for its activity. Each of the six Cys residues in human PNMT was changed to Ser by PCR-based site-directed mutagenesis, and each mutant PNMT was expressed in Escherichia coli to identify the functionally important Cys residue. The six mutants (C48S, C60S, C91S, C131S, C139S, and C183S) and the wild-type enzyme were expressed at almost the same levels as revealed by Western blotting analysis. Kinetic parameters (apparent Km and Vmax) of C48S, C60S, C91S, C131S, and C139S for the substrates, norepinephrine and S-adenosyl-L-methionine, showed similar values to those of the wild-type enzyme. However, C183S exhibited markedly reduced enzyme activity with less than 3% of the wild-type Vmax and with ca. sixfold increased apparent Km values for both substrates. These results suggested that Cys183 plays an important role in the activity of human PNMT.

[Chronic fatigue syndrome and psychiatric diseases].

The chronic fatigue syndrome consists of a combination of non-specific symptoms. Some believe that the CFS is subcategory of major depression, because the symptoms are similar to those of major depression. We believe that the CFS is quite different from major depression or neurotic depression, since the CFS has no lack of initiative and effort, no inhibition which is seen in endogenous depression, and sharp fluctuations in general fatigue, anxiety, and various persisting somatic symptoms, such as, malaise and mild fever. CFS seems to be similar to the neurasthenia. It is harmful, at least, in aetiology and treatment, to neglect the diagnosis of the CFS.

Radiation-induced osteosarcoma of the sphenoid bone.

The case of a patient who developed osteosarcoma in the sphenoid bone 15 years after radiation therapy for a craniopharyngioma is reported. Radiation-induced osteosarcoma of the sphenoid bone has not been reported previously. Reported cases of radiation-induced osteosarcomas are reviewed.

Scrapie-associated fibrils (SAF) purification method yields amyloid proteins from systemic and cerebral amyloidosis.

We identified fibrils from non-transmissible systemic and cerebral amyloidosis using the purification method of scrapie-associated fibrils (SAF). The fibrils possessed the same nature of congophilia, filamentous structures and molecular weights as amyloid fibrils, and were resistant to Proteinase K digestion. This SAF method makes for a rapid extraction from amyloid-laden tissues. The method, therefore, may purify nontransmissible amyloids alone or together with SAF proteins.

Re-evaluation of protein-bound glutathione in rat liver.

The extent of intracellular glutathione binding to proteins through a disulfide linkage in rat liver was examined quantitatively. The content of glutathione associated with the acid-precipitable fraction and releasable on borohydride treatment was 0.024 +/- 0.016 mumol/g liver, which accounted for less than one per cent of the total glutathione (6-7 mumol/g liver) in the liver of fed rats. Most of the thiol (2-4 mumol/g liver) liberated from liver proteins into the acid-soluble fraction on borohydride reduction in the presence of guanidine hydrochloride was not glutathione but was proteinaceous in nature. The amounts of thiols liberated per g of liver were similar in fed, fasted, and dibutyryl-3',5'-cyclic AMP-treated rats.

Morphogenesis of amyloid plaques in mice with Creutzfeldt-Jakob disease.

Amyloid plaques were experimentally produced in the brains of mice inoculated with human Creutzfeldt-Jakob disease (CJD) brain homogenate. Light-microscopically, these plaques were mostly round and oval and were surrounded by macrophages and astrocytes. Ultramicroscopically, amyloid plaques were present in the cytoplasm of macrophages or were surrounded by these cells. The macrophages had numerous Golgi apparatuses, endoplasmic reticula (ER), ribosomes, polysomes, and lysosomes with inoculated materials or degenerating products. The bundles of amyloid fibrils were intermingled with the cytoplasm of macrophages, and sometimes limiting membranes were absent. Some bundles of amyloid fibrils projected from the Golgi apparatuses or rough ER and were partly exposed to the extracellular spaces, but there were no amyloid fibrils in the lysosomes. These findings confirmed that amyloid fibrils in the brains of CJD-infected mice were produced by macrophages.

A new method to classify amyloid fibril proteins.

The permanganate method, the immunoperoxidase method, and a newly developed autoclave method were used to distinguish different types of amyloid fibril proteins in formalin-fixed, paraffin-embedded tissue sections. All tissues from permanganate-sensitive cases (AA type) lost the affinity of Congo red and green birefringence under polarized light after incubation with special autoclave treatment. AL type systemic amyloidosis and amyloid plaques of CJD and GSS were permanganate-resistant, but decreased markedly the affinity of Congo red after prolonged autoclaving. On the other hand, prealbumin type systemic amyloidosis and senile plaques of SDAT were resistant to both permanganate oxidation and prolonged autoclaving. Thus, amyloid plaques of CJD and GSS are identical to AL type in systemic amyloidosis, and senile plaques are similar to the prealbumin type. However, anti-prealbumin antiserum did not stain senile plaque amyloid. The anti-human P component stained positively systemic amyloids and cerebral amyloid plaques of SSE, but failed to stain senile plaques of SDAT. Therefore, the amyloid fibril protein of senile plaques is apparently different from other types of amyloid depositions. Amyloid plaques of SSE are different from senile plaques not only with regard to fibril proteins, but also to globular protein in the amyloid.

Amyloid plaques in the brains of mice with Creutzfeldt-Jakob disease.

By intracerebral inoculation with the brain homogenates from 9 patients with Creutzfeldt-Jakob disease, amyloid plaques were induced in mouse brains after incubation periods of 403 to 835 days. The plaques existed mainly in the cerebral white matter beneath the lateral ventricle walls and were more numerous in the hemisphere where injection was made. Morphological findings of the plaques were almost identical to those seen in patients with kuru and Creutzfeldt-Jakob disease, and in animals with scrapie. They were also similar to the cores of senile plaques seen in patients with Alzheimer's disease.